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Item does not contain fulltext BACKGROUND: Rezafungin is a next-generation, once-a-week echinocandin in development for the treatment of candidaemia and invasive candidiasis and for the prevention of invasive fungal disease caused by Candida, Aspergillus, and Pneumocystis spp after blood and marrow transplantation. We aimed to compare the efficacy and safety of intravenous rezafungin versus intravenous caspofungin in patients with candidaemia and invasive candidiasis. METHODS: ReSTORE was a multicentre, double-blind, double-dummy, randomised phase 3 trial done at 66 tertiary care centres in 15 countries. Adults (≥18 years) with systemic signs and mycological confirmation of candidaemia or invasive candidiasis were eligible for inclusion and randomly assigned (1:1) to receive intravenous rezafungin once a week (400 mg in week 1, followed by 200 mg weekly, for a total of two to four doses) or intravenous caspofungin (70 mg loading dose on day 1, followed by 50 mg daily) for no more than 4 weeks. The primary endpoints were global cure (consisting of clinical cure, radiological cure, and mycological eradication) at day 14 for the European Medical Agency (EMA) and 30-day all-cause mortality for the US Food and Drug Administration (FDA), both with a target non-inferiority margin of 20%, assessed in the modified intention-to-treat population (all patients who received one or more doses of study drug and had documented Candida infection based on a culture from blood or another normally sterile site obtained within 96 h before randomisation). Safety was evaluated by the incidence and type of adverse events and deaths in the safety population, defined as all patients who received any amount of study drug. The trial is registered with ClinicalTrials.gov, NCT03667690, and is complete. FINDINGS: Between Oct 12, 2018, and Aug 29, 2021, 222 patients were screened for inclusion, and 199 patients (118 [59%] men; 81 [41%] women; mean age 61 years [SD 15·2]) were randomly assigned (100 [50%] patients to the rezafungin group and 99 [50%] patients to the caspofungin group). 55 (59%) of 93 patients in the rezafungin group and 57 (61%) of 94 patients in the caspofungin group had a global cure at day 14 (weighted treatment difference -1·1% [95% CI -14·9 to 12·7]; EMA primary endpoint). 22 (24%) of 93 patients in the rezafungin group and 20 (21%) of 94 patients in the caspofungin group died or had an unknown survival status at day 30 (treatment difference 2·4% [95% CI -9·7 to 14·4]; FDA primary endpoint). In the safety analysis, 89 (91%) of 98 patients in the rezafungin group and 83 (85%) of 98 patients in the caspofungin group had at least one treatment-emergent adverse event. The most common treatment-emergent adverse events that occurred in at least 5% of patients in either group were pyrexia, hypokalaemia, pneumonia, septic shock, and anaemia. 55 (56%) patients in the rezafungin group and 52 (53%) patients in the caspofungin group had serious adverse events. INTERPRETATION: Our data show that rezafungin was non-inferior to caspofungin for the primary endpoints of day-14 global cure (EMA) and 30-day all-cause mortality (FDA). Efficacy in the initial days of treatment warrants evaluation. There were no concerning trends in treatment-emergent or serious adverse events. These phase 3 results show the efficacy and safety of rezafungin and support its ongoing development. FUNDING: Cidara Therapeutics and Mundipharma.

Antimicrobial stewardship (AMS) is a coherent set of actions to optimize antimicrobial use, improve patient outcomes, and reduce antimicrobial resistance (AMR). Thailand's national strategic plan has included AMS since 2017. Due to an increased risk of infection and AMR, solid organ transplant (SOT) recipients can benefit from antimicrobial stewardship programs (ASPs). However, the AMS in SOT has not been well defined. Balancing ASPs with the need for early and frequent antimicrobial treatment in this population is challenging. This review assesses the barriers and potential strategies of AMS in SOT in the setting of Thailand.We used PubMed to identify published articles on AMS in organ transplantation in Thailand from January 2013 to January 2022. We also searched local literature and local data from Google Scholar and Google. Finally, we described the AMS experience at Ramathibodi hospital as a proxy for transplant centers in Thailand.There was no specific article on AMS in SOT in Thailand. At our hospital, ASPs have been part of the hospital's routine patient care, including the practice in the transplant unit. Modifiable challenges to AMS in SOT include prescriber opposition, diagnosis uncertainty, and lack of high quality. Both systems and individualized approaches should be implemented to overcome these barriers. The potential interventions include shortening antibiotic time-out, updating clinical guidelines, continuing education, handshake ASP, adopting new technology, and further research.There are limited data on AMS in organ transplantation in Thailand. Strategic priorities should focus on modifiable barriers tailored to organ transplantation. Quality improvement should be ensured by process and outcome measures.

Purpose The incidence of carbapenem-resistant Enterobacteriaceae (CRE) has been increasing worldwide. Ertapenem resistance is mediated by non-carbapenemase mechanisms, and has less of an effect on susceptibility to imipenem and meropenem. This study aimed to study the epidemiology of CRE, and to compare risk factors and related mortality between non-susceptibility to ertapenem alone Enterobacteriaceae (NSEE), with non-susceptibility to other carbapenems (imipenem, meropenem, or doripenem) Enterobacteriaceae (NSOCE) at a tertiary care hospital in Thailand. Methods All CRE isolated were identified between December 2011 and December 2016. Quarterly incidence rate was estimated. Hospital-wide carbapenem consumption was calculated as defined daily doses (DDD). Relationships between hospital-wide carbapenem consumption and incidence of CRE were tested. Factors associated with NSEE and NSOCE, and risk factors associated with 14- and 30-day mortality in patients with CRE infection were determined. Results The quarterly CRE incidence increased significantly from 3.37 per 100,000 patient-days in the last quarter of 2011 to 32.49 per 100,000 patient-days in the last quarter of 2016. (P for trend

Background Reusing N95 respirators with a practical and easy decontamination protocol might be a solution for the shortage of N95 respirators. We aimed to study the reliability and safety of household UV-C devices for the decontamination of N95 respirators. Methods We investigated a baby bottle ultraviolet C (UV-C) sterilizer box for N95 decontamination. Swine coronavirus (PEDV) was sprayed on the N95 and the virus was cultivated after UV-C sterilization. Bacterial culture from used N95 respirator was performed before and after UV-C exposure. Scanning electron Microscopy (SEM) was used to observe the structural change after UV-C exposure. The radiation was measured by radiometer. The fit test was performed on 3 participants before use, and after reuse 4 and 6 times. Results The PEDV sprayed on the N95 respirator was inactivated by the UV-C level of 0.930–0.932 mW/cm2. Nearly all bacterial colonies had disappeared after the 10 min of the UV-C exposure. No significant change to the structure of the N95 polymer fibers after 240 min of UV-C exposure was observed by SEM. Three and two participants passed the fit test after the fourth and sixth time of reuse, respectively. Conclusions During the COVID-19 pandemic crisis, UV-C products may be practical and safe options for decontaminating N95 respirators, if the energy of UV-C is in the range known to deactivate the virus.

BackgroundPCR is more sensitive than immunofluorescence assay (IFA) for detection ofPneumocystis jirovecii. However, PCR cannot always distinguish infection from colonization. This study aimed to compare the performance of real-time PCR and IFA for diagnosis ofP.jiroveciipneumonia (PJP) in a real-world clinical setting.MethodsA retrospective cohort study was conducted at a 1,300-bed hospital between April 2017 and December 2018. Patients whose respiratory sample (bronchoalveolar lavage or sputum) were tested by bothPneumocystisPCR and IFA were included. Diagnosis of PJP was classified based on multicomponent criteria. Sensitivity, specificity, 95% confidence intervals (CI), and Cohen's kappa coefficient were calculated.ResultsThere were 222 eligible patients. The sensitivity and specificity of PCR was 91.9% (95% CI, 84.0%–96.7%) and 89.7% (95% CI, 83.3%–94.3%), respectively. The sensitivity and specificity of IFA was 7.0% (95% CI, 2.6%–14.6%) and 99.2% (95% CI, 95.6%–100.0%), respectively. The percent agreement between PCR and IFA was 56.7% (Cohen's kappa -0.02). Among discordant PCR-positive and IFA-negative samples, 78% were collected after PJP treatment. Clinical management would have changed in 14% of patients using diagnostic information, mainly based on PCR results.ConclusionsPCR is highly sensitive compared with IFA for detection of PJP. Combining clinical, and radiological features with PCR is useful for diagnosis of PJP, particularly when respiratory specimens cannot be promptly collected before initiation of PJP treatment.

Objectives The aim of this study was to determine the incidence and predictive factors of catheter-related bloodstream infections (CRBSIs) acquired through a central venous catheter for delivery of home parenteral nutrition (HPN) therapy among a patient population in Thailand. Methods This retrospective review was conducted with adult patients with intestinal failure who received HPN from October 2002 to April 2014, at Ramathibodi Home Parenteral and Enteral Nutrition Referral Center in Bangkok. Results Seventy-two patients (45.8% male, mean age 56.2 ± 15.7 y) were included in the analysis. Incidence of CRBSIs was 1.47 per 1000 catheter days. Over the 12-y period, there were 21 CRBSIs among 10 patients. There were 26 pathogens isolated from these CRBSIs, mostly coagulase-negative staphylococci. Univariate Poisson regression analysis revealed that the incidence rate ratio of CRBSIs was significantly higher in patients who used an implanted port (compared with tunneled catheter), alcohol-based povidone–iodine solution as disinfectant (compared with 2% chlorhexidine gluconate in 70% isopropyl alcohol), cyclic HPN infusion (compared with continuous HPN), and hospital-based compound HPN formulations (compared with 3-in-1 commercial formulations). Furthermore, longer duration of HPN (>250 d) was associated with CRBSIs. Multivariate analysis revealed that longer duration of HPN, cyclic HPN, and hospital-based compound HPN were significantly associated with CRBSIs. Conclusions CRBSI is a significant complication in patients receiving long-term HPN. Individualized therapy with a multidisciplinary team in centers with HPN management expertise is required. Careful selection of the catheter type and HPN formulation for each patient is necessary to best meet patient requirements and minimize HPN-related complications. Strict compliance by patients and caregivers with evidence-based instructions together with supervision by well-trained HPN providers is the most effective strategy to prevent CRBSIs.

OBJECTIVESThe role of methicillin-resistant Staphylococcus aureus (MRSA) nasal surveillance swabs (nasal swabs) in guiding decisions about prescribing vancomycin is unclear. We aimed to determine the likelihood that patients with negative MRSA nasal swabs develop subsequent MRSA infections; to assess avoidable vancomycin days for patients with negative nasal swabs; and to identify risk factors for having a negative nasal swab and developing a MRSA infection during the intensive care unit (ICU) stay.METHODSThis retrospective cohort study was conducted in 6 ICUs at a tertiary-care hospital from December 2013 through June 2015. The negative predictive value (NPV), defined as the ability of a negative nasal swab to predict no subsequent MRSA infection, was calculated. Days of vancomycin continued or restarted after 3 days from the collection time of the first negative nasal swab were determined. A matched case-control study identified risk factors for having a negative nasal swab and developing MRSA infection.RESULTSOf 11,441 patients with MRSA-negative nasal swabs, the rate of subsequent MRSA infection was 0.22%. A negative nasal swab had a NPV of 99.4% (95% confidence interval [CI], 99.1%–99.6%). Vancomycin was continued or started after nasal swab results were available in 1,431 patients, translating to 7,364 vancomycin days. No risk factors associated with MRSA infection were identified.CONCLUSIONSIn our hospital with a low prevalence of MRSA transmission, a negative MRSA nasal swab was helpful in identifying patients with low risk of MRSA infection in whom empiric vancomycin therapy could be stopped and in whom the subsequent initiation of vancomycin therapy during an ICU admission could be avoided.Infect Control Hosp Epidemiol 2018;39:290–296

Background The recommended duration of antibiotic treatment for Enterobacteriaceae bloodstream infections is 7-14 days. We compared the outcomes of patients receiving short-course (6-10 days) vs prolonged-course (11-16 days) antibiotic therapy for Enterobacteriaceae bacteremia. Methods A retrospective cohort study was conducted at 3 medical centers and included patients with monomicrobial Enterobacteriaceae bacteremia treated with in vitro active therapy in the range of 6-16 days between 2008 and 2014. 1:1 nearest neighbor propensity score matching without replacement was performed prior to regression analysis to estimate the risk of all-cause mortality within 30 days after the end of antibiotic treatment comparing patients in the 2 treatment groups. Secondary outcomes included recurrent bloodstream infections, Clostridium difficile infections (CDI), and the emergence of multidrug-resistant gram-negative (MDRGN) bacteria, all within 30 days after the end of antibiotic therapy. Results There were 385 well-balanced matched pairs. The median duration of therapy in the short-course group and prolonged-course group was 8 days (interquartile range [IQR], 7-9 days) and 15 days (IQR, 13-15 days), respectively. No difference in mortality between the treatment groups was observed (adjusted hazard ratio [aHR], 1.00; 95% confidence interval [CI], .62-1.63). The odds of recurrent bloodstream infections and CDI were also similar. There was a trend toward a protective effect of short-course antibiotic therapy on the emergence of MDRGN bacteria (odds ratio, 0.59; 95% CI, .32-1.09; P = .09). Conclusions Short courses of antibiotic therapy yield similar clinical outcomes as prolonged courses of antibiotic therapy for Enterobacteriaceae bacteremia, and may protect against subsequent MDRGN bacteria.

Background Carbapenem-resistant Enterobacteriaceae (CRE) has been increasing worldwide. Our objectives were to study the epidemiology of CRE and compare risk factors and mortality of carbapenem nonsusceptibility to ertapenem alone Enterobacteriaceae (NSEE) with nonsusceptibility to other carbapenems (imipenem, meropenem, or doripenem) Enterobacteriaceae (NSOCE) at a tertiary care hospital in Thailand. Methods All CRE isolated from clinical and surveillance cultures were identified from December 2011 to December 2016. Quarterly incidence rate per 100,000 patient-days was estimated. Hospital-wide carbapenem consumption were calculated as defined daily doses (DDD) per 1,000 patient-days. Relationships between hospital-wide carbapenem consumption and incidence of CRE were tested using Poisson regression. Comparative analysis of factors associated with NSEE and NSOCE, and risk factors associated with 14- and 30-day mortality in patients with CRE infection was conducted in adult patients. Results The quarterly CRE incidence of unique patients increased significantly from 3.37 per 100,000 patient-days in the last quarter of 2011 to 32.49 per 100,000 patient-days in the last quarter of 2016. Quarterly CRE incidence increased 1.07 per 100,000 patient-days (95% confidence interval [CI], 0.49–1.06; P-value for trend Conclusion The incidence of CRE has risen significantly over a 5-year period at our institution. The important risk factor for nonsusceptibility to other carbapenems compared with nonsusceptibility to ertapenem alone was previous carbapenem use. Our hospital-wide carbapenem use has significantly increased over time, and associated with the increasing incidence of CRE. Disclosures All authors: No reported disclosures.

This retrospective study described the first reported vancomycin-resistant enterococci (VRE) outbreak from June 2013 through January 2014 at a tertiary-care hospital in Bangkok, Thailand. After the index case was detected in an 18-bed medical intermediate care unit, a number of interventions was implemented, including targeted active surveillance for VRE, strict contact precautions, enhanced standard precautions, dedicated units for VRE cases, extensive cleaning of the environment and the restricted use of antibiotics. VRE isolates were evaluated by polymerase chain reaction and random amplified polymorphic DNA (RAPD) testing. A prevalence case-control study was conducted. Among 3,699 culture samples from 2,671 patients screened, 74 patients (2.8%) had VRE. The positivity rate declined from 15.1% during week 1 to 8.2% during week 2 and then 1.4% during week 3. By weeks 4-9, the prevalences were 0-2.7%. However, the prevalence rose to 9.4% during week 10 and then subsequently declined. All VRE isolates were Enterococcus faecium and had the vanA gene. RAPD analysis revealed a single predominant clone. Multivariate analysis showed mechanical ventilation for ≥ 7 days was a predictive factor for VRE colonization [odds ratio (OR) 11.47; 95% confidence interval (CI): 1.75-75.35; p = 0.011]. This experience demonstrates VRE can easily spread and result in an outbreak in multiple-bed units. Active surveillance, early infection control interventions and rapid patient cohorting were important tools for control of this outbreak. Patients requiring mechanical ventilator for ≥ 7 days were at higher risk for VRE acquisition.

In this prospective cohort study, we investigated the prevalence of Clostridium difficile-associated diarrhea (CDAD) in adult patients with nosocomial diarrhea by performing enzyme immunoassay (EIA) for detecting toxins A and B and polymerase chain reaction (PCR) for detecting the presence of the tcdB gene in stool samples. We determined the factors associated with CDAD, and the treatment outcome of CDAD from May 2010 to January 2011. A total of 175 stool samples were tested by EIA and PCR. In total, 26.9% patients tested positive for C. difficile: 12.6% by EIA and 24.0% by PCR. The kappa coefficient and total agreement of both the tests were 0.46 and 83.2%, respectively. Onset of diarrhea after antibiotic administration for 10 days or more (OR, 2.71; 95% CI, 1.14-6.44; P = 0.024) and leukocyte count >15,000 cells/mm(3) (OR, 3.12; 95% CI, 1.24-7.88; P = 0.016) were significantly associated with occurrence of CDAD. The non-response rate to CDAD treatment was 24.1%, and the all-cause mortality rate was 31.9% in the CDAD group as against 35.9% in the non-CDAD group (P = 0.721). In our study, the performance of direct PCR of stool samples for detecting tcdB was better, with the number of positive results for stool toxins A and B being twofold higher than that in the case of EIA. Patients who have diarrhea after receiving antibiotics for 10 days or more or those who have a leukocyte count of >15,000 cells/mm(3) should be investigated for CDAD.

Although hepatitis B serology screening has been recommended for HIV care, it has not been routinely performed. We aimed to assess compliance and timing of hepatitis B serology screening among HIV-infected patients in a resource-limited setting. A cross-sectional study was conducted in Thailand. Compliance, timing of hepatitis B serology screening, and factors associated with no HBsAg screening were determined. A total of 416 HIV-infected patients with 61% males were enrolled. Median (range) age at HIV diagnosis was 34 (16-75) years and 92% had heterosexual risk. Proportion of HBsAg screening and prevalence of positive HBsAg were 69.2% and 9.0%, respectively. There was no difference in the proportion of no HBsAg screening during the period 1990-2008 (p = 0.865). Proportion of anti-HBs and anti-HBc screening were 40.9% and 21.2%, respectively. HBsAg was screened before or on the day of anti-HIV testing in 9.1% and before antiretroviral therapy (ART) initiation in 27.2%. By Kaplan-Meier analysis, median time from anti-HIV testing to HBsAg screening was 55.9 (95% confidence interval [CI] 43.9, 68.3) months. By multivariate logistic regression, duration of HIV infection (odds ratio [OR] 1.14; 95% CI 1.07, 1.21), no anti-HBs screening (OR 1.65; 95% CI 1.4-2.63), and no anti-HCV screening (OR 2.60; 95% CI 1.62, 4.17) were associated with no HBsAg screening before ART initiation. In conclusion, compliance with hepatitis B serology screening was relatively low and late. Educational program regarding hepatitis B serology screening, identification of barriers, and interventions to eliminate these barriers in resource-limited settings are crucial to improve HIV care.

Background The recommended duration of antibiotic treatment for Enterobacteriaceae bacteremia is between 7 and 14 days. We compared the clinical outcomes of patients receiving short-course (6–10 days) vs prolonged-course (11–15 days) antibiotic therapy for Enterobacteriaceae bacteremia. Methods A retrospective cohort study was conducted at The Johns Hopkins Hospital, The University of Maryland Medical Center, and The Hospital of the University of Pennsylvania including patients with monomicrobial Enterobacteriaceae bacteremia treated with in vitro active antibiotic therapy in the range of 6–15 days between 2008 and 2014. 1:1 nearest neighbor propensity score matching without replacement was performed, prior to regression analysis, to estimate the risk of all-cause mortality within 30 days after the end of antibiotic treatment for patients receiving short vs. prolonged durations of antibiotic therapy. Secondary outcomes included Clostridium difficile infection (CDI) and the emergence of multidrug-resistant Gram-negative (MDRGN) bacteria within 30 days after the end of antibiotic therapy. Results A total of 1,769 patients met eligibility criteria. There were 385 matched pairs who were well-balanced on baseline characteristics. The median duration of therapy in the short-course group and prolonged-course group was 8 days (interquartile range (IQR) 7–9 days) and 15 days (IQR 13–15 days), respectively. No difference in all-cause mortality between short- and prolonged-course treatment groups was observed (adjusted hazard ratio [aHR] 1.00; 95% CI 0.62–1.63). Rates of CDI were similar between the treatment groups (OR 1.17; 95% CI 0.39–3.51). There was a non-significant protective effect of short-course antibiotic therapy on the emergence of MDRGN bacteria (OR 0.59; 95% CI 0.32–1.09 P = 0.09). Conclusion Short courses of antibiotic therapy yields similar clinical outcomes to prolonged courses of antibiotic therapy for Enterobacteriaceae bacteremia, and may protect against subsequent MDRGN emergence. Disclosures All authors: No reported disclosures.

Background The role of MRSA nasal surveillance swabs in guiding decisions about need for subsequent vancomycin therapy is unclear. Our objectives were to (1) determine the likelihood that patients with negative MRSA nasal swabs went on to develop MRSA infections during the same hospitalizations to assess if vancomycin therapy could be avoided once the nasal swab result returns negative, (2) assess days of vancomycin that potentially could be avoided, and (3) identify risk factors for having a negative MRSA nasal swab and developing an MRSA infection during the hospital stay. Methods This retrospective cohort study was conducted at six intensive care units (ICUs) at a tertiary care hospital in Baltimore from December 2013 to June 2015. MRSA nasal swabs are obtained at the time of admission and weekly thereafter for all ICU patients. The negative predictive value (NPV), defined as the ability of a negative MRSA nasal screening test to correctly predict no subsequent MRSA infection during the hospital stay, was calculated, accounting for the 3-day turnaround time of MRSA nasal surveillance swabs. Days of vancomycin therapy started or continued after 3 days from the first negative MRSA nasal swab were determined by chart review. A matched case–control study was performed to identify risk factors for patients with negative MRSA surveillance cultures who subsequently developed MRSA infections. Results Of 11,441 MRSA-nasal swab negative patients, the proportion of subsequent incident MRSA infections was 0.2%. Negative MRSA surveillance swabs had an NPV of 99.4% (95% CI 99.1–99.6%). Among 4,091 MRSA-negative patients receiving vancomycin, vancomycin was started or continued after 3 days since the first MRSA-negative nasal swab in 1,434 patients (35%), translating to 7,377 potentially avoidable vancomycin days. The matched case–control analysis did not identify risk factors associated with subsequent MRSA infection. Conclusion At our institution with robust infection control practices and low nosocomial MRSA transmission rates, patients with negative MRSA nasal swabs have a very low likelihood of subsequent MRSA infection during hospitalizations. MRSA nasal swabs can provide useful information when determining whether to initiate or stop empiric vancomycin. Disclosures All authors: No reported disclosures.

We report the case of a 64-year-old man with Arcanobacterium pyogenes endocarditis. The patient presented with dyspnea and asymmetrical progressive quadriparesis. A transthoracic echocardiogram revealed mobile vegetations on both leaflets of his mitral valve measuring 0.5 x 3 cm, thickening of the mitral valve with severe mitral regurgitation due to dehiscence of the papillary muscle to the posterior mitral leaflet. He also had aortic sclerosis with a vegetation measuring 0.5 x 1 cm causing aortic valve dehiscence and free flow aortic regurgitation. An initial hemoculture grew out pleomorphic, gram-positive, non-motile, anaerobic to microaerophilic bacilli. A diagnosis of infective endocarditis was made using modified Duke criteria. He was treated with intravenous ampicillin and gentamicin. Four days after admission, he developed acute respiratory failure and succumbed to the disease. A pre-mortem hemoculture and post-mortem heart valve culture grew Arcanobacterium pyogenes. Septic thromboemboli involving the brain, kidneys, lungs and spleen were documented. The patient also had ischemic vasculopathy with focal spinal arteriolitis and bilateral demyelination of the cervical corticospinal tracts. There are three published reports of human A. pyogenes endocarditis in the literature. Neurological involvement with ischemic spinal vasculopathy and demyelination has not been reported. We report the first autopsy proven case of A. pyogenes infective endocarditis with ischemic spinal vasculopathy. We review the clinicopathologic features of systemic A. pyogenes infection.

Clostridium difficileposes as the most common etiologic agent of nosocomial diarrhea. Although there are many diagnostic methods to detectC. difficiledirectly from stool samples, the nucleic acid-based approach has been largely performed in several laboratories due to its high sensitivity and specificity as well as rapid turnaround time. In this study, a multiplex PCR was newly designed with recent accumulated nucleotide sequences. The PCR testing with variousC. difficileribotypes, otherClostridiumspp., and non-Clostridiumstrains revealed 100% specificity with the ability to detect as low as ~22 genomic copy number per PCR reaction. Different combinations of sample processing were evaluated prior to multiplex PCR for the detection ofC. difficilein fecal samples from hospitalized patients. The most optimal condition was the non-selective enrichment at 37∘C for 1 h in brain heart infusion broth supplemented with taurocholate, followed by the multiplex PCR. The detection limit after sample processing was shown as being 5 spores per gram of fecal sample. Two hundred and thirty-eight fecal samples collected from the University affiliated hospital were analyzed by the enrichment multiplex PCR procedure. The results suggested that the combination of sample processing with the high-performance detection method would be applicable for routine diagnostic use in clinical setting.